Nano fluorescent probe, and preparation method and application thereof in detection of HNO in Golgi apparatus

A nano-fluorescent probe, fluorescein technology, used in fluorescence/phosphorescence, chemical instruments and methods, luminescent materials, etc., can solve the problems of short existence time, limitation, difficult capture, etc., and achieve low phototoxicity, cell and living body damage. The effect of small, simple synthetic route

Active Publication Date: 2020-03-17
SHANDONG NORMAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, because HNO is a substance that can react spontaneously, it is extremely difficult to capture in vivo for a short time. Therefore, the methods and means of direct detection of HNO still need to be further developed, which also limits the ability of HNO in many physiological and pathological processes in living cells and in vivo environments. Conducting research on effects

Method used

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  • Nano fluorescent probe, and preparation method and application thereof in detection of HNO in Golgi apparatus
  • Nano fluorescent probe, and preparation method and application thereof in detection of HNO in Golgi apparatus
  • Nano fluorescent probe, and preparation method and application thereof in detection of HNO in Golgi apparatus

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0045] Synthesis of fluorescent probes

[0046] Dissolve the raw material 2-diphenylphosphobenzoic acid (5mmol), dicyclohexylcarbodiimide (5mmol), and 4-dimethylaminopyridine (0.25mmol) in 15mL of dichloromethane, activate the carboxyl group at 0°C for 30min, Fluorescein (1 mmol) was then added and stirred at room temperature for 12 h. After the reaction was complete, the solvent was removed by rotary evaporation. Then using dichloromethane:methanol=20:1 as eluent, purified by column chromatography to obtain yellow solid C-HNO (57%).

[0047]Dissolve 2 mg of C-HNO in 300 μL of dimethyl sulfoxide, quickly add it into 100 mg of BSA previously dissolved in 10 mL of deionized water with a pipette, and stir at room temperature for 1 hour. After the reaction was stopped, dialyze overnight with a dialysis bag with a molecular weight of 3000 to obtain an aqueous solution of nano fluorescent probe BSA-HNO with a concentration of 100 μg / mL.

[0048] Effect experiment:

[0049] Gener...

Embodiment 2

[0056] Synthesis of fluorescent probes

[0057] Dissolve the raw materials 2-diphenylphosphobenzoic acid (1mmol), dicyclohexylcarbodiimide (1mmol), and 4-dimethylaminopyridine (0.05mmol) in 15mL of dichloromethane, activate the carboxyl group at 0°C for 30min, Fluorescein (1 mmol) was then added and stirred at room temperature for 12 h. After the reaction was complete, the solvent was removed by rotary evaporation. Then using dichloromethane:methanol=20:1 as eluent, purified by column chromatography to obtain yellow solid C-HNO (15%).

[0058] Dissolve 2 mg of C-HNO in 300 μL of dimethyl sulfoxide, quickly add it into 100 mg of BSA previously dissolved in 10 mL of deionized water with a pipette, and stir at room temperature for 1 hour. After the reaction was stopped, dialyze overnight with a dialysis bag with a molecular weight of 3000 to obtain an aqueous solution of nano fluorescent probe BSA-HNO with a concentration of 100 μg / mL.

Embodiment 3

[0060] Synthesis of fluorescent probes

[0061] Dissolve the raw material 2-diphenylphosphobenzoic acid (3mmol), dicyclohexylcarbodiimide (3mmol), and 4-dimethylaminopyridine (0.15mmol) in 15mL of dichloromethane, activate the carboxyl group at 0°C for 30min, Fluorescein (1 mmol) was then added and stirred at room temperature for 12 h. After the reaction was complete, the solvent was removed by rotary evaporation. Then using dichloromethane:methanol=20:1 as eluent, purified by column chromatography to obtain yellow solid C-HNO (24%).

[0062] Dissolve 2 mg of C-HNO in 300 μL of dimethyl sulfoxide, quickly add it into 100 mg of BSA previously dissolved in 10 mL of deionized water with a pipette, and stir at room temperature for 1 hour. After the reaction was stopped, dialyze overnight with a dialysis bag with a molecular weight of 3000 to obtain an aqueous solution of nano fluorescent probe BSA-HNO with a concentration of 100 μg / mL.

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Abstract

The invention provides a nano fluorescent probe, and a preparation method and an application thereof in detection of HNO in a Golgi apparatus. The nano fluorescent probe comprises a probe molecule C-HNO and bovine serum albumin (BSA), the bovine serum albumin coats the probe molecule C-HNO, and the chemical structural formula of the probe molecule C-HNO is shown in the specification. The nano fluorescent probe shows an excellent Golgi apparatus targeting positioning effect, can be used for imaging HNO, and has the advantages of high sensitivity, high selectivity and simplicity and conveniencein synthesis.

Description

technical field [0001] The disclosure relates to a Golgi body target positioning detection technology, in particular to a nano fluorescent probe and a preparation method thereof and its application in detecting HNO in the Golgi body. Background technique [0002] The information disclosed in this Background section is only intended to increase the understanding of the general background of the disclosure, and is not necessarily to be taken as an acknowledgment or any form of suggestion that the information constitutes prior art that is already known to those skilled in the art. [0003] The Golgi apparatus is a secretory subcellular organelle composed of many flat membrane-structured vesicles. Golgi apparatus exists widely in eukaryotic cells and is closely related to many physiological and pathological processes. Its main functions include cooperating with the endoplasmic reticulum to synthesize proteins, and further processing, sorting and transport. Proteins processed a...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C09K11/02C09K11/06A61K49/00C07F9/6561G01N21/64
CPCC09K11/025C07F9/6561C09K11/06G01N21/6428A61K49/0021A61K49/0056C09K2211/1007C09K2211/1014C09K2211/1088G01N2021/6417
Inventor 唐波刘翠芳张晓婷王慧李平
Owner SHANDONG NORMAL UNIV
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